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Literature summary for 3.5.1.4 extracted from

  • Silman, N.J.; Carver, A.M.; Jones, C.W.
    Directed evolution of amidase in Methylophilus methylotrophus; purification and properties of amidases from wild-type and mutants strains (1991), J. Gen. Microbiol., 137, 169-178.
No PubMed abstract available

Inhibitors

Inhibitors Comment Organism Structure
DTNB 15-18% inhibition of wild-type, MM6 and MM8, 35% inhibition of the MM15 activity Methylophilus methylotrophus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2
-
acetamide mutant strain MM8 Methylophilus methylotrophus
2
-
Acrylamide mutant strain MM8 Methylophilus methylotrophus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
38000
-
4 * 38000 Methylophilus methylotrophus
38400
-
6 * 38400 Pseudomonas aeruginosa
40000
-
8 * 40000 Arthrobacter sp.
155000
-
sedimentation-equilibrium ultracentrifugation Methylophilus methylotrophus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acetamide + H2O Methylophilus methylotrophus
-
acetic acid + NH3
-
?
acylamide + H2O Methylophilus methylotrophus
-
carboxylic acid + NH3
-
?

Organism

Organism UniProt Comment Textmining
Arthrobacter sp.
-
-
-
Methylophilus methylotrophus
-
wild-type is strain MM6, mutant strains are MM8 and MM15
-
Pseudomonas aeruginosa
-
-
-

Purification (Commentary)

Purification (Comment) Organism
passage with French pressure cell, cell-free extract purified and concentrated by using anion-exchange and gel filtration, judged by SDS-PAGE and scanning densiometry Methylophilus methylotrophus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.51
-
mutant strain MM15 measured with acrylamide as substrate, growth conditions is continuous Methylophilus methylotrophus
2.1
-
mutant strain MM15 measured with acetamide as substrate Methylophilus methylotrophus
3.59
-
mutant strain MM15 measured with acrylamide as substrate, growth conditions is fed-batch Methylophilus methylotrophus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetamide + H2O
-
Methylophilus methylotrophus acetic acid + NH3
-
?
acrylamide + H2O
-
Methylophilus methylotrophus acrylate + NH3
-
?
acylamide + H2O
-
Methylophilus methylotrophus carboxylic acid + NH3
-
?

Subunits

Subunits Comment Organism
hexamer 6 * 38400 Pseudomonas aeruginosa
octamer 8 * 40000 Arthrobacter sp.
tetramer 4 * 38000 Methylophilus methylotrophus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.13
-
acetamide
-
Methylophilus methylotrophus